zebrafish embryo protein isolation

FLUET at BIOMED.MED.YALE.EDU FLUET at BIOMED.MED.YALE.EDU
Fri Feb 24 11:59:31 EST 1995


Hello all,

I have been using the protocol from The Zebrafish Book to extract proteins 
from zebrafish embryos (around 28 h or so) for Western analysis (page 9.2). 
 I've had good luck with it, but have not been able to quantify the amount of 
proteins with the Bio-Rad (Bradford) protein assay kit.  I think the 3.5% 
SDS in the sample buffer is the culprit.  Has anyone tried another 
detergent, like 1% NP-40 or maybe Triton X-100?  Also, I thought I might 
use DTT instead of beta-Mercaptoethanol. Any advice/comments you all might 
have would be appreciated.  Thanks in advance.


Amy Fluet 




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